Advance Journal of Food Science and Technology 6(5): 616-621, 2014

DOI:10.19026/ajfst.6.84
ISSN: 2042-4868; e-ISSN: 2042-4876
© 2014 Maxwell Scientific Publication Corp.
Submitted: January 16, 2014 Accepted: January 25, 2014 Published: May 10, 2014

Research Article
Purification of Sardine Oil Using Adsorbent (Active Filter) of Scallop Shells,
Carp Scales and Attapulgite
Sugeng Heri Suseno, Yosephina M.J. Batafor, Nurjanah and Ayu Fitri Izaki
Department of Aquatic Product Technology, Faculty of Fisheries and Marine Sciences, Bogor
Agricultural University, Jl. Agatis Darmaga 16680 Bogor, Indonesia

Abstract: Fish oil especially from sardine has contain great omega 3, but it needs purification because has poor
quality. Fish oil purification can be done by using various types of adsorbents. One of them are scallop shells and
carp scales. The purpose of this study is to choose the best type of active filter (adsorbent) to improve the quality of
fish oil. Sardine oil is purified using an active filter (adsorbent) of carp scales, scallop shells and attapulgite. The
result this study show that the best treatment of adsorbent types is carp scales+scallop shell adsorbent.

Keywords: Active filter, adsorbent, refining, sardine oil, secondary primary oxidation

INTRODUCTION adsorbate through direct contact surface. Additionally

Sardine (Sardinella lemuru) has great potential as a
source of fish oil (5-20 %) and omega 3. Fish oil is
useful for health because it contains a lot of unsaturated
fatty acids with many double bonds (PUFA) omega 3
that is Eicosapentaenoic acid (EPA) and Docosa
Hexaenoic Acid (DHA) are beneficial to the body
(Aidos, 2002). Omega 3 fatty acids are most abundant
in fish oil are EPA, AA and DHA. These fatty acids can
cure various diseases such as atherosclerosis (narrowing
and hardening of the arteries), thrombosis mellitus and
other joint or bone disease, asthma and prevent the
aging process (Duthie and Barlow, 1992).
Fishmeal and canning process of sardine produces
by-product fish oils containing large enough omega 3
fatty acids in high levels and can be used as a source of
omega 3 fatty acids. Fish oil purification efforts with an
effective and efficient method are very important to
improve the quality of fish oils so suitable for
consumption. One is the method of active filtration.
Active filtration serves to fix the color of the oil,
reducing unwanted odor components, sulfur
compounds, heavy metals and can reduce the
production of fat oxidation are peroxides, aldehydes
and ketones (Estiasih, 2009). Fish oil purification can
be carried out by making use of various kinds of
adsorbents. Scallop shells and carp scales are one of the
by-product of the fishery that has not been properly
utilized and wasted into waste. Esmaeli et al. (2012)
and Checa et al. (2007) showed that the results of
Scanning Electron Microscope (SEM) on the structure
of carp scales and shells of the bivalve have extensive
porous surface that can be used to physically adsorb an
scallop shells and carp scales containing chitin that
serve as metal ion chelating. Treatment of active
filtration is done by using a material that has the ability
to adsorption impurity components. The purpose of this
study is to choose the best type of active filter
(adsorbent) to improve the quality of fish oil.
MATERIALS AND METHODS
The main materials used in this study are crude
sardine oil from Bali industry. The adsorbent used was
carp scales (Cyprinus carpio) obtained from Bogor
market, scallop shells (Amusium pleuronectes) came
from Banten and attapulgite obtained from the
Laboratory of Pharmacy, University of Pancasila. Carp
scales and scallop shells sun dried and destroyed so that
the form of flour, while attapulgite has been shaped
flour. Other supporting materials are the materials used
for the analysis of the quality of fish oil in the form of
distilled water, glacial acetic acid, chloroform, a
solution of potassium iodine (KI) saturated solution of
sodium thiosulfate (Na2S2O3) 0.1 N, 0.1 N KOH
solution, the indicator of phenolphthalein, 96 ethanol,
1% starch, isooktan, reagent p-anisidine, n-hexane. The
equipments used for the purification of fish oil include
erlenmeyer glass, aluminum foil, magnetic stirrer,
digital scales, pipette and high speed refrigerated
centrifuge himac brand HITACHI CR 21G. Other
support equipment such as buret, stative, 2500 UV-Vis
spectrophotometer LaboMed brands, bulbs, test tubes
and pumpkin drinks.

Corresponding Author: Sugeng Heri Suseno, Department of Aquatic Product Technology, Faculty of Fisheries and Marine
Sciences, Bogor Agricultural University, Jl. Agatis Darmaga 16680 Bogor, Indonesia
This work is licensed under a Creative Commons Attribution 4.0 International License (URL: http://creativecommons.org/licenses/by/4.0/).
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 Adv. J. Food Sci. Technol., 6(5): 616-621, 2014

Fish oil purification: Sardine oil is purified using an

active filter (adsorbent) of carp scales, scallop shells
and attapulgite. There are 12 treatment used that is
control (C), carp scales (CS), scallop shells (SS),
attapulgite (A), carp scales + scallop shells (CS + SS),
carp scales + attapulgite (CS + A), scallop shell +
attapulgite (CS + A), carp scales + scallop shells +
attapulgite (CS + SS + A), gradually treatment carp
scales and scallop shells (CS-SS), gradually treatment
carp scales and attapulgite (CS-A), gradually treatment
scallop shells and attapulgite (SS-A), gradually
treatment carp scales, scallop shells and attapulgite
(CS-SS-A), 3% adsorbent concentration, agitation time
fish oil and adsorbent using magnetic stirrer for 20 min
at room temperature (±29°C), after which a mixture of
fish oil and adsorbent, then separated by centrifuging at Fig. 1: Free fatty acid value of sardine oil; Note: CS (carp
a speed of 10000 rpm for 30 min at a temperature of scales Cyprinus carpio), SS (scallop shells Amusium
10°C. pleuronectes) and A (attapulgite). sign 'plus'
(combined addition of adsorbent), sign 'arrow'
Analysis of fish oil quality: Oil purifying results using (gradual addition of adsorbent)
the active filter (adsorbent) then analyzed free fatty acid
value (AOCS, 1998, method No. Ca 5a-40), peroxide
value (AOAC, 2000, method No. 965.33b), p-anisidine
value (IUPAC, 1987, method No. 2.504), total
oxidation value (AOCS, 1997) and clarity (wavelength
450, 550, 620, 665, 700 nm, respectively) (AOAC,
1995).

Data analysis: Result of purifying process was

statistically processed using ANOVA by SPSS software
version 16.0 to see the regression parameter
coefficients, percent significance (confidence interval)
and the pattern of interaction of factors that
significantly influence the response.

RESULTS AND DISCUSSION
Free fatty acid (FFA): Analysis of free fatty acids is
an analysis conducted to determine the amount of free
fatty acids formed by the breakdown of oil through the
chemical process of hydrolysis and biologically by
microbes. The results of the analysis of free fatty acids
of sardine oils with types of adsorbent treatment are
presented in Fig. 1.
The results of the analysis of the value of free fatty
acids sardine oil showed the highest value generated by
gradually treatment of scallop shell and attapulgite
adsorbent (12.48±0.63%) and the lowest value in the
treatment of carp scales+shell scallop adsorbent
(6.70±0.07%). Statistical analysis showed that the type
of adsorbent treatment affected (p<0.05) to the value of
free fatty acids (Fig. 1). Bimbo (1998) suggest
specification quality crude fish oil with a free fatty acid
content ranged from 1 to 7%, but typically ranges from
2 to 5%, while Young (1986) states that the maximum
acceptable value of 4.0%. The presence of free fatty
acids in oil due to oil undergoes hydrolysis process.
Hydrolysis of triglycerides in the oil will produce fatty
acids and monoglycerides components (Winarno,
2004). The difference decreased levels of free fatty acid
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Fig. 2: Peroxide value of sardine oil; Note: CS (carp scales
Cyprinus carpio), SS (scallop shells Amusium
pleuronectes) and A (attapulgite). sign (gradual
addition of adsorbent)
levels may occur by the differences in the type of
adsorbent used. Different types of adsorbents will have
a polarity, the active surface, surface area, porosity,
particle size, pH and different of water content (Zhu
et al., 1994).
Peroxide Value (PV): Peroxide value is the most
important value to determine the degree of damage to
the oil or fat. The results of the analysis of the peroxide
value of sardine oil with treatment of adsorbent types
are presented in Fig. 2.
The results of the analysis of the peroxide value of
sardine oil showed the highest value generated by
gradually treatment of carp scale and scallop shell
adsorbent (14.25±1.25 meq/kg) and the lowest value in
the treatment of carp scales + scallop shells adsorbent
(6.00±0.50 meq/kg). Statistical analysis showed that the
type of adsorbent treatment affected (p<0.05) on
peroxide value (Fig. 2). Peroxide value with the type of
 Adv. J. Food Sci. Technol., 6(5): 616-621, 2014

adsorbent treatment not meets the standards of the

International Fish Oil Standard (IFOS (International
Fish Oils Standard), 2011) (≤3.75 meq/kg). Standard
peroxide value according to the Council for
Responsible Nutrition (CRN (Council for Responsible
Nutrition), 2006) is 5 meq/kg. Increased peroxide is
because each stage of the process requires the
characterization of the oil temperature and long storage
time. Boran et al. (2006) reported the storage conditions
of fish oil and fatty acid profiles contained in fish oil
affect oxidative damage. The quality of the fish used for
oil extraction, oil extraction process and storage
conditions will affect peroxide value of crude fish oil
(EFSA (European Food Safety Authority), 2010). Fig. 3: P-anisidine value of sardine oil; Note: CS (carp scales
Winarno (2004) oil containing unsaturated fatty acids Cyprinus carpio), SS (scallop shells Amusium
are high susceptible to oxidation. Indicating a high pleuronectes) and A (attapulgite). sign 'plus'
peroxide value of fat or oil has been oxidized, but at a (combined addition of adsorbent), sign 'arrow'
lower value is not always indicate that the oxidation (gradual addition of adsorbent)
state is still early.

P-anisidine value (P-AV): P-anisidine value analysis is

an analysis that is performed to measure secondary
oxidation products (carbon component) (AOCS, 1994).
The results of the analysis of the p-anisidine value of
sardine oil with adsorbent types of treatment are
presented in Fig. 3.
The results of the analysis of the p-anisidine value
of sardine oil showed the highest value generated by
treatment of carp scales+scallop shells adsorbent
(0.26±0.05 meq/kg) and the lowest value in the
treatment of scallop shells adsorbent (0.22±0.15
meq/kg). Statistical analysis showed that the type of
adsorbent treatment did not affect (p>0.05) on p-
anisidine value (Fig. 3). P-anisidine value with
treatment type adsorbents can meet the standards of the
International Fish Oil Standard (IFOS (International
Fish Oils Standard), 2011) (≤15 meq/kg). P-anisidine
value is performed to measure the carbon components
that influence the formation of unpleasant odors (off-
flavor) results from the oxidation process (EFSA
(European Food Safety Authority), 2010).
Total oxidation value (Totox): Analysis of total
oxidation is an analysis conducted to determine the
presence of different compounds such as
hydroperoxides, aldehydes, ketones, mainly produced
by the degradation of PUFA in pro-oxidant conditions,
especially high temperature, oxygen and light metal
compounds. The results of the analysis of the total
oxidation of sardine oil with adsorbent types of
treatment are presented in Fig. 4.
The results of the analysis of the total oxidation
value of sardine oil showed the highest value generated
by treatment of carp scales and scallop shells adsorbent
(28.75±2.46 meq/kg) and the lowest value in the
treatment of carp scales+scallop shells adsorbent
(12.26±1.05 meq/kg). Statistical analysis showed that
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Fig. 4: Total oxidation value of sardine oil; Note: CS (carp
scales Cyprinus carpio), SS (scallop shells Amusium
pleuronectes) and A (attapulgite). sign (gradual
addition of adsorbent)
the type of adsorbent treatment affected (p<0.05) to the
total oxidation value (Fig. 4). The best total oxidation
value of the treatment that is adsorbent types of carp
scales+scallop shells (12.26±1.05 meq/kg) to meet the
standards of the International Fish Oil Standards (IFOS
(International Fish Oils Standard), 2011) (≤20 meq/kg).
The total oxidation value is calculated of two times the
peroxide value plus the value of the p-anisidine. It is
used to measure the total oxidation of oil by
considering both primary and secondary products of
oxidation, can be used for assessment of fish oil
oxidation during storage (CRN (Council for
Responsible Nutrition), 2006). The pore size is quite
important role in the adsorption process. Vitara (2007)
states that molecules with large size are difficult to get
into the pores or voids contained in the adsorbent pore
size are smaller when compared to the molecule.
Factors affecting of the adsorption capacity which is the
surface area, pore size, solubility of the adsorbate, pH
 Adv. J. Food Sci. Technol., 6(5): 616-621, 2014

(e)
(a)
Fig. 5: Percent clarity of fish oil transmission after the
addition adsorbent with a wavelength of 450 (a), 550
(b), 620 (c), 665 (d) and 700 nm (e)

and temperature. The main constituent of the shells of

shrimp or shellfish is chitin, a natural polysaccharide
which has many uses such as a chelating agent,
emulsifier and adsorbent (Bhuvana, 2006).

Clarity value: Measurement of fish oil clarity

performed at 5 wavelengths, i.e., 450, 550, 620, 665
and 700 nm. Fish oil purity level is indicated by the
value of percent transmission read on a
spectrophotometer. Percent value of high transmission
(b) and approaching 100% indicates that fish oil was
observed to have a good level of clarity. The results of
the analysis of percent light transmission to the sardine
oil with the treatment type of adsorbent at 5
wavelengths are presented in Fig. 5.
The results of the analysis of fish oil percent light
transmission at a wavelength of 450 nm showed the
highest value generated by treatment of carp scales
adsorbent (60.30±16.50%) and the lowest value in the
treatment of attapulgite adsorbent (41.50±0.30%).
Statistical analysis showed that the type of adsorbent
treatment affected (p<0.05) to the value of clarity (Fig.
5a). Figure 5a shows at a wavelength of 450 nm, carp
scales are the most effective adsorbent to purify fish oil
(c) seen from the greatest percent transmission.
The results of the analysis of fish oil percent light
transmission at a wavelength of 550 nm showed the
highest value generated by gradually treatment of carp
scales, scallop shell and attapulgite (89.55±2.05%) and
the lowest value in the treatment of carp scales
adsorbent (78.35±5:05%). Statistical analysis showed
that the type of adsorbent treatment affect (p<0.05) to
the value of clarity (Fig. 5b). Figure 5b shows at a
wavelength of 550 nm, carp scales scallop and shell-
attapulgite is the most effective adsorbent to purify fish
oil seen from the greatest percent transmission.
The results of the analysis of fish oil percent light
transmission at a wavelength of 620 nm showed the
(d) highest value generated by gradually treatment of
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 Adv. J. Food Sci. Technol., 6(5): 616-621, 2014
scallop shell and attapulgite adsorbent (98.00±1.30%)
and the lowest value in the treatment of attapulgite
adsorbent (91.35±0.75%). Statistical analysis showed
that the type of adsorbent treatment affect (p<0.05) to
the value of clarity (Fig. 5c). Figure 5c shows at a
wavelength of 620 nm, attapulgitescallop shell is the
most effective adsorbent to purify fish oil seen from the
greatest percent transmission.
The results of the analysis of fish oil percent light
transmission at a wavelength of 665 nm showed the
highest value generated by treatment of carp scales+
scallop shells adsorbent (98.45±1.15%) and the lowest
value in the treatment of attapulgite adsorbent
(93.45±0.75%). Statistical analysis showed that the type
of adsorbent treatment affected (p<0.05) to the value of
clarity (Fig. 5d). Figure 5d shows at a wavelength of
665 nm, carp scales+scallop shell is the most effective
adsorbent to purify fish oil seen from the greatest
percent transmission.
The results of the analysis of fish oil percent light
transmission at a wavelength of 700 nm showed the
highest value generated by the treatment of adsorbent
carp scales+scallop shells (99.65±0:15%) and the
lowest value in the treatment of attapulgite adsorbent
(96.75±0.65%). Statistical analysis showed that the type
of adsorbent treatment affected (p<0.05) to the value of
clarity (Fig. 5d). Figure 5d shows at a wavelength of
700 nm, carp scales+scallop shell is the most effective
adsorbent to purify fish oil seen from the greatest
percent transmission.
CONCLUSION
Fish oil purification can be done by using
adsorbent such as carp scales, scallop shells and
attapulgite. Carp scales+scallop shell is the best
adsorbent treatment characteristics, seen from the total
oxidation that can meet the standards of IFOS
(International Fish Oils Standard) (2011). The best
value with clarity percent light transmission values
approaching 100 scales contained in the adsorbent
treatment carp scales+scallop shells at a wavelength of
665 nm and 700 nm.
ACKNOWLEDGMENT
This study completion would not have been
possible without the assistance of many people who
gave their support. To them I would like to convey my
heartfelt gratitude and sincere appreciation.
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